The structure and the effect of polysaccharide from peach gum (DPG2) on ameliorating DSS-induced acute colitis in mice were investigated in the present study. The results showed that DPG2 was identified as an AG II arabinogalactan with the backbone of β-D-(1 → 6)-galactan, which consisted of mannose, glucuronic acid, galactose, xylose and arabinose with a molar ratio of 4.64:1.02:2.61:39.82:3.89:48.02. Moreover, DPG2 behaved as a flexible chain conformation with a coil-like structure with a molecular weight (Mw) of 5.21 × 105 g/mol. Furthermore, the worm-like chain model parameters for DPG2 were estimated as follows: ML = 379 nm-1, q = 0.74 nm and d = 0.82 nm. The results of the animal assay showed that the intake of DPG2 not only effectively improved the phenotypes of DSS-induced colitis in mice, but also significantly improved the oxidative stress status of mice, such as regulating NO content and T-SOD and MPO levels and repairing oxidative damage to the colonic mucosa. Moreover, DPG2 improved the inflammation of DSS-induced colitis in mice by inhibiting the secretion of the proinflammatory cytokines TNF-α, IFN-γ, IL-1β, IL-6 and IL-17. Therefore, these results suggested that peach gum polysaccharide showed protective effects against colitis, and has great potential for the application of functional components in the food industry.